A DeltaVision fluorescence microscopy system (Applied Precision), which is based on an Olympus wide-field IX71 fluorescence microscope equipped with an oil-immersion objective lens (Plan Apo 60×; NA = 1.4; Olympus) and CoolSNAP HQ2 CCD camera (Photometrics), was used to image the yeast cells. For time-lapse observation, living cells were mounted on 35-mm glass-bottomed culture dishes (MatTek) coated with 0.2 mg/mL soybean lectin (Sigma) and observed at 26 °C unless otherwise specified. A set of images of 15 focal planes at 0.5 μm intervals was taken at each time point. Images were deconvolved using the DeltaVision SoftWorx software (Applied Precision).
Deltavision softworx software
Manufactured by Cytiva
Sourced in United States
DeltaVision softWoRx software is a comprehensive imaging and analysis suite designed to work with Cytiva's DeltaVision microscopy systems. The software provides tools for image acquisition, processing, and analysis, enabling users to capture, visualize, and quantify biological samples with high resolution and accuracy.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Merck Group (7 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (6 mentions)
Co2 independent media, by Thermo Fisher Scientific (4 mentions)
Glass bottom culture dishes, by MatTek (4 mentions)
Spinning disk confocal microscope, by PerkinElmer (2 mentions)
Soybean lectin, by Merck Group (1 mentions)
Planapo 60, by Olympus (1 mentions)
Deltavision fluorescence microscopy system, by Cytiva (1 mentions)
35 mm glass bottomed culture dishes, by MatTek (1 mentions)
Softworx, by Cytiva (1 mentions)
Deltavision microscopy system, by Cytiva (1 mentions)
Anti α tubulin, by Merck Group (1 mentions)
Anti hif 1α, by Cell Signaling Technology (1 mentions)
12 protocols using deltavision softworx software
1
Fluorescence Microscopy of Yeast Cells
2
Localization of KmYME in K. marxianus
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Localization of KmYME in K. marxianus was conducted as described previously (Mo et al., 2016 (link)). Imaging was conducted using a Perkin Elmer spinning-disk confocal microscope (PerkinElmer, Inc., Norwalk, CT, United States) equipped with a Zeiss PlanApo 100X/1.4 NA objective and a Photometrics EMCCD camera Evolve 51234. Images were captured by the DeltaVision softWoRx software (Applied Precision) and processed by deconvolution and z-stack projection. All images were analyzed using ImageJ.
3
Immunofluorescence and Time-lapse Imaging of HeLa Cells
4
Immunofluorescence Staining of MDA-MB-231 Cells
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MDA-MB-231 cells grown on coverslips were fixed using PBS buffer supplemented with 3.7% paraformaldehyde. After blocking with PBS with 0.05% Tween-20 (PBST) buffer containing 1% bovine serum albumin (Sigma) for 45 min at room temperature, the fixed cells were incubated with primary antibodies in a humidified chamber for 1 h at room temperature, followed by secondary antibodies for 1 h. The DNA was stained with 4′,6-diamidino-2-phenylindole from Sigma. Images were captured by DeltaVision softWoRx software (Applied Precision) and processed by deconvolution and z-stack projection.
5
Immunofluorescence and Time-lapse Imaging of HeLa Cells
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HeLa cells grown on coverslips were fixed by a pre-extraction method using PTEM buffer (60 mM PIPES, pH7.9; 10 mM EGTA; 2 mM MgCl2; 0.2% Triton X-100) supplemented with 3.7% paraformaldehyde. After blocking with PBST (PBS with 0.05% Tween-20) buffer containing 1% bovine serum albumin (Sigma) for 45 min at room temperature, the fixed cells were incubated with primary antibodies in a humidified chamber for 1 h at room temperature or overnight at 4 °C, followed by secondary antibodies for 1 h at 37 °C. The DNA was stained with 4′,6-diamidino-2-phenylindole (DAPI) from Sigma. Images were captured by DeltaVision softWoRx software (Applied Precision) and processed by deconvolution and z-stack projection.
For Time-lapse imaging, HeLa cells were cultured in glass-bottom culture dishes (MatTek) and maintained in CO2-independent media (Gibco) supplemented with 10% FBS and 2 mM glutamine49 (link). During imaging, the dishes were placed in a sealed chamber at 37 °C. Images of living cells were taken with a DeltaVision microscopy system.
For Time-lapse imaging, HeLa cells were cultured in glass-bottom culture dishes (MatTek) and maintained in CO2-independent media (Gibco) supplemented with 10% FBS and 2 mM glutamine49 (link). During imaging, the dishes were placed in a sealed chamber at 37 °C. Images of living cells were taken with a DeltaVision microscopy system.
6
Immunofluorescence Imaging of CT26 Cells
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The procedure
for CT26 cell culture is the same as that for the western blot. Following
different treatments, cells were fixed with 4% paraformaldehyde (PFA)
at 37 °C for 20 min and permeabilized with PBS containing 0.2%
Triton X-100 at 37 °C for 1 min. After blocking with PBS containing
0.05% Tween-20 buffer and 5% bovine serum albumin (Sigma) at room
temperature for 120 min, the cells were incubated with primary antibodies
against HIF-1α or calreticulin or PD-L1 in a humidified chamber
at room temperature for 120 min, followed by phalloidin (Rhodamine
Conjugate) at room temperature for 2 h. Images were captured by DeltaVision
SoftWoRx Software (Applied Precision) and processed by deconvolution
and z-stack projection.
for CT26 cell culture is the same as that for the western blot. Following
different treatments, cells were fixed with 4% paraformaldehyde (PFA)
at 37 °C for 20 min and permeabilized with PBS containing 0.2%
Triton X-100 at 37 °C for 1 min. After blocking with PBS containing
0.05% Tween-20 buffer and 5% bovine serum albumin (Sigma) at room
temperature for 120 min, the cells were incubated with primary antibodies
against HIF-1α or calreticulin or PD-L1 in a humidified chamber
at room temperature for 120 min, followed by phalloidin (Rhodamine
Conjugate) at room temperature for 2 h. Images were captured by DeltaVision
SoftWoRx Software (Applied Precision) and processed by deconvolution
and z-stack projection.
7
Immunofluorescence and Live-cell Imaging of Mog1 in HeLa Cells
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HeLa cells transfected with Mog1 siRNA or mCerulean-Mog1 were fixed using PTEM buffer (60 mM PIPES, pH6.8, 10 mM EGTA, 2 mM MgCl2, 0.2% Triton X-100) supplemented with 3.7% paraformaldehyde. After blocking with 1% bovine serum albumin (Sigma-Aldrich) in PBST (PBS with 0.05% Tween-20) buffer for 45 min at room temperature, the fixed cells were incubated with primary antibodies in a humidified chamber for 1 h followed by secondary antibodies for 1 h at room temperature. The DNA was stained with DAPI (Sigma-Aldrich). Images were acquired by DeltaVision SoftWoRx software (Applied Precision) and processed by deconvolution and z-stack projection.
For live cell imaging, HeLa cells were cultured in glass-bottom culture dishes (MatTek) and maintained in CO2-independent media (Gibco) supplemented with 10% (v/v) FBS and 2 mM glutamine (Ding et al., 2010 (link)). During imaging, the dishes were placed in a sealed chamber at 37°C. Images of living cells were taken with a DeltaVision microscopy system (Applied Precision Inc.). Image processing was performed with SoftWoRx (Applied Precision Inc.). To trace chromosomes in mitosis, frames were collected at 3–5 min intervals. Images were prepared for publication using Adobe Photoshop software.
For live cell imaging, HeLa cells were cultured in glass-bottom culture dishes (MatTek) and maintained in CO2-independent media (Gibco) supplemented with 10% (v/v) FBS and 2 mM glutamine (Ding et al., 2010 (link)). During imaging, the dishes were placed in a sealed chamber at 37°C. Images of living cells were taken with a DeltaVision microscopy system (Applied Precision Inc.). Image processing was performed with SoftWoRx (Applied Precision Inc.). To trace chromosomes in mitosis, frames were collected at 3–5 min intervals. Images were prepared for publication using Adobe Photoshop software.
8
Immunofluorescence and Time-lapse Imaging of HeLa Cells
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For immunofluorescence, HeLa cells were plated onto a sterile 24-well plate (Corning Glass Works) for transfection or drug treatment and fixed with 3.7% formaldehyde in PBS for 10 min at 37°C. In order to facilitate the staining of the antibody into the cells, the treated and fixed cells were permeated with PBS containing 0.1% Triton X-100 for 5 min. After blocking with PBS with 0.05% Tween-20 buffer containing 1% bovine serum albumin (Sigma) for 45 min at room temperature, the fixed cells were incubated with primary antibodies in a humidified chamber for 1 h at room temperature or overnight at 4°C, followed by secondary antibodies for 1 h at 37°C. The DNA was stained with 4′,6-diamidino-2-phenylindole from Sigma. Images were captured by DeltaVision softWoRx software (Applied Precision) and processed by deconvolution and z-stack projection.
For time-lapse imaging, HeLa cells were cultured in glass-bottom culture dishes (MatTek) and maintained in CO2-independent media (Gibco) supplemented with 10% FBS and 2 mM glutamine (Mo et al., 2016 (link)). During imaging, the dishes were placed in a sealed chamber at 37°C. Images of living cells were taken with a DeltaVision microscopy system.
For time-lapse imaging, HeLa cells were cultured in glass-bottom culture dishes (MatTek) and maintained in CO2-independent media (Gibco) supplemented with 10% FBS and 2 mM glutamine (Mo et al., 2016 (link)). During imaging, the dishes were placed in a sealed chamber at 37°C. Images of living cells were taken with a DeltaVision microscopy system.
9
Immunolabeling and Imaging of Neurons
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HEK293 cells were immunolabeled in live and fixed/permeabilized conditions [13 (link)]. Slides were imaged using an inverted confocal microscope (63X 1.4 numerical aperture oil immersion lens) (Leica, Germany) and analyzed using Image J v1.46 software (National Institutes of Health, Bethesda, Maryland). Embryonic day 16.5 mouse hippocampal neurons were cultured as previously described [13 (link), 17 (link)]. Immunocytochemistry on live primary murine neurons and human neural stem cell-derived neurons were performed as previously described [13 (link), 41 (link)]. Neurons were visualized through 100X 1.4 numerical aperture oil immersion lens with an inverted Olympus IX-70 microscope (DeltaVision Core, Applied Precision, GE Healthcare, Washington) and a CoolSnap QE camera (Photometrics, Tucson, Arizona). Images were acquired as 0.15 μm-thick 40 serial optical sections, then deconvolved using DeltaVision SoftWoRx software, version 5.0.0 (Applied Precision, GE Healthcare), and volume projections of the entire Z-series were generated and overlaid using ImageJ. All procedures on animals were approved by the Kids Research Institute and Children’s Medical Research Institute animal ethics committee and conformed to the published code of practice of the National Health and Medical Research Council of Australia.
10
Immunofluorescence Imaging of HIF-1α in 4T1 Cells
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4T1 cells were seeded in confocal dish. Following different treatments, cells were fixed with 1% paraformaldehyde at 37 °C for 10 min and permeabilized with PBS containing 0.2% Triton X-100 at 37 °C for 1 min. Subsequently, the blocking step was carried out with PBST (PBS with 0.05% Tween-20) buffer containing 1% bovine serum albumin at room temperature for 45 min. Finally, the cells were incubated with anti-HIF-1α (Cat. no. 36169T, Cell Signaling Technology) and anti-α-Tubulin (Cat. no. T9026, Sigma-Aldrich) primary antibodies in a humidified chamber at 4 °C overnight, followed by corresponding fluorescence labeled secondary antibodies at room temperature for 1 h. Images were acquired with a DeltaVision softWoRx software (Applied Precision) and processed by deconvolution and z-stack projection.
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